Related Product Information

Introduction

Dynabeads Pan Mouse IgG in combination with primary mouse IgG antibodies are ideal for depletion or positive isolation of cells from different species (e.g. human, rat) depending on the specificity of the primary antibody. Cells can be directly isolated from any sample such as whole blood, bone marrow, MNC suspensions or tissue digests.

The primary mouse IgG antibody is either added to the cell sample (indirect technique) or pre-coated onto the beads (direct technique) prior to cell isolation. Dynabeads are then mixed with the cell sample in a tube. The Dynabeads will bind to the target cells during a short incubation, and then the beadbound cells are separated by a magnet.

Positive isolation – discard the supernatant and use the bead-bound cells for downstream applications (e.g. molecular analysis or cell culture).

Depletion – discard the bead-bound cells and use the remaining, untouched cells for any application.

Use a mixer that provides tilting and rotation of the tubes to ensure Dynabeads do not settle at the bottom of the tube. When incubating Dynabeads and cells, the incubation temperature must be 2-8°C to reduce phagocytic activity and other metabolic processes. Never use less than 25 μl (1 x 107) Dynabeads per ml cell sample and at least 4 Dynabeads per target cell.

Table 1: Volume of Dynabeads added per ml of cell sample. The volumes can be scaled up as required.

Positive isolation

Depletion

Sample volume (1 x 107 cells/ml*)

1 ml

1 ml

Volume of Dynabeads

25 μl

50 μl

Total no. of cells processed per product

2 x 109 cells

1 x 109 cells

* If the concentration of cells is increased, the Dynabeads volume must be increased accordingly. Cell concentration can be up to 1 x 108cells per ml

Cell Isolation - Indirect Technique

Labeling Cells with Mouse IgG Antibodies

Use approximately 1 μg of primary antibody (mouse IgG) per 106 target cells.

The choice of primary antibody is the most important factor for successful cell isolation. Note that some antibodies may show reduced antigen binding efficiency when coated onto beads (direct technique), even though the antibody shows good results in other immunological assays.

Remove soluble factors in serum: Serum may contain soluble factors (e.g. antibodies or cell surface antigens), which can interfere with the cell isolation protocol. Washing the cells once may reduce this interference.

General Information

Invitrogen Dynal AS complies with the Quality System Standards ISO 9001:2000 and ISO 13485:2003.

Storage/Stability

This product is stable until the expiry date stated on the label when stored unopened at 2-8°C. Store opened vials at 2-8°C and avoid bacterial contamination. Keep Dynabeads in liquid suspension during storage and all handling steps, as drying will result in reduced performance. Resuspend well before use.

Warnings And Limitations

This product is for research use only. Not intended for any animal or human therapeutic or diagnostic use unless otherwise stated. Follow appropriate laboratory guidelines. This product contains 0.02% sodium azide as a preservative, which is cytotoxic.

Avoid pipetting by mouth!

Sodium azide may react with lead and copper plumbing to form highly explosive metal azides. When disposing through plumbing drains, flush with large volumes of water to prevent azide build up. Certificate of Analysis (CoA) is available upon request. Material Safety Data Sheet (MSDS) is available at
.